CTMJ | traditionalmedicinejournals.com                                    Chinese Traditional Medicine Journal | 2021 | Vol4 |Issue3 

 
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 Chinese Traditional Medical Journal  

Pharmacological Research 

Antimicrobial Activity of Vitex Negundo Linn. (Nirgundi) Leaves 

Extract 

Samijk Ullahg, Liu Leiley 

Department of Botany, University of Peshawar, Peshawa 

, Department of Graduate School of Tianjin University of Traditional Chinese Medicine, Tianjin China 

 

 

 

 

 

 

 

 

 

 

 

 

 

Introduction 

icrobes, such as viruses, bacteria, and parasites, 

are all around us, some of which are helpful to 

our health and others which are detrimental. 

Infectious illnesses account for 26% of all 

fatalities each year and over 30% of all DALYs 

(disability-adjusted life years) lost each year. [1] 

More than three million children die each year 

from malaria and diarrheal illnesses alone in 

impoverished nations. [2] Antibiotics are the 

medicines that suppress or kill germs. 

Developing novel medications to treat 

Abstract 
Background: Antibiotic resistance in human pathogens has been more widespread during the last 
several years. This necessitates the development of antibacterial medications that are not based on 
antibiotics. Aims: in vitro testing of the antibacterial properties on both f and g gramme of Vitex 
negundo Linn. (Nirgundi) organisms with a gramme positive and a gramme negative stain 
 Material and Methods: To test the antimicrobial activity of Vitex negundo Linn. leaf oethanol 
extracts against three strains of bacteria obtained from the American Type Culture Collection (ATCC 
25922, ATCC 25923, and ATCC 27736), researchers used the well diffusion technique (ATCC). The 
concentrations of 20 mg/ml, 40 mg/ml, and 60 mg/ml were used in this investigation. Leaf extract 
concentrations of up to 100 mg/ml were utilised. 
 Results: All of the bacterial pathogens investigated were shown to have antibacterial properties. 
Highest ZOI recorded for S. aureus was 15 mm at 80 and 100 mg/ml, respectively, whereas the 
maximum ZOI was 12 mm for E. coli and 11 mm for K. pneumoniae at 100 mg/ml for the other two 
pathogens. 
 Conclusion: A significant antibacterial effect was found for S aureus in the Vitex negundo Linn. leaf 
extract when compared to all the other microorganisms. 
  
Keywords: Antimicrobial Property, Vitex negundo Linn., Nirgundi, Well Diffusion Method 

 



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infections is necessary since bacteria are 

capable of transmitting and acquiring resistance 

to pharmaceuticals used as therapeutic agents 

[3]. 

Bacteria appear often in ancient works of 

literature. Vedic literature has identified and 

described over a hundred different species of 

microorganisms. There are macroscopic and 

microscopic creatures (like Drishya) in the world 

(Adrishya). They might come from inside or 

outside the body. They've located the main 

(Vesa) and secondary (Parivesha) locations of 

infection in the body. The organisms should be 

wiped off in order to lessen their negative 

impacts. For example, herbal medications such 

as Manidharana (wearing jewels), Bali 

(sacrificing an animal), Homa (votive ritual), and 

Mantra (hymns) were used, as well as the sun, 

re, and Guggulu. [4] Sankramaka or infectious 

disorders were identified by Sushrut and 

Vagbhata. Some of these disorders have been 

linked to microbes, according to the research. 

Because of this, antimicrobial medications 

known as Krumighna or Rakshoghna have been 

described in Ayurvedic texts for centuries. [5] 

[6] 

As a member of the verbenaceae family, Vitex 

negundo grows to a height of up to 4.5 metres 

and is distributed across India. Therapeutic 

properties include anti-inflammatories, 

antibiotics, antibacterials, moderate CNS 

depressants antifertility, analgesic, hepato-

protective and estrogenic anticonvulsant. 

Herbalists employ the leaves for anything from 

rheumatism and arthritis to catarrhal fever and 

syphilis to sprains and inflamed sores and 

ulcers. [7] Antibiotic-resistant pathogens, as 

well as their adverse effects, might be harmful 

to the host. As a result of the pressing demand 

for novel antimicrobial medications with fewer 

side effects than synthetic pharmaceuticals, the 

current research examined the antibacterial 

activity of Nirgundi leaves. Results were 

collected at Jawaharlal Nehru Medical College 

in Wardha, Maharashtra, India, in the 

department of microbiology.. 

Plant material: T From the 21st of July to the 

30th of September of 2014, fresh leaves of Vitex 

negundo Linn. were harvested in the hamlet of 

Dabha, Wardha District (MS), which is located 

at 20.57 N and 78.81 E. Herbarium was made 

and delivered to the Botanical Survey of India in 

Pune, Maharashtra, for plant certification. No. 

BSI/WRC-Technology/2014/447-MSD-4 is a 

voucher specimen. 

 

Washing leaves under tap water for 2-3 times, 

followed by an ethanol wash, and then allowing 

them to dry in the shade at room temperature 

for seven days removes soil and other 

contaminants. Using a mixer grinder, we 

pulverised the leaves of three different plants 

until we had a coarse powder. Polyethylene 

bags containing the powder were secured, 

marked, and placed in an airtight container for 

further investigation. 

Preparation of extracts: A soxhlet device was 

used to extract 20 gm of powdered Vitex 

negundo Linn. leaves into 200 ml of ethanol at 

40-500C. 2.29 gramme of the filtered extract 

was collected and stored at room temperature 

to eliminate any remaining ethanol. And 10 cc 

of ethanol was used to dilute the solution. The 

mother solution was kept in a refrigerator for 

future usage and was referred to as 20 mg/ml, 

40 mg/ml, 60 mg/ml, 80 mg/ml, and 100 mg/ml 

concentrations of the extract. 

Microorganisms: E. coli, Staphylococcus aureus 

and Klebsiella pneumoniae bacteria were 



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purchased from the American Type Culture 

Collection (ATCC) for use in this study (ATCC). 

Preparation of subculture media for Inoculum: 

The agar slant culture was transferred into the 

sterilised nutrient broth medium through a 

loopful of organism. At 370 degrees Fahrenheit, 

the tubes were incubated for 18-24 hours. 

Preparation of Assay Medium and Pour plates: 

After sterilising the Petri plates, Mueller Hinton 

Agar (MHA) was poured into them and allowed 

to cool at 400C until it solidified. There is a lot 

of room for  

 

 

 

development and culture in the Petri dishes. A 

sterile swab steak was used to mow the 

prepared subculture media (a combination of 

microorganisms) over the surface of the solid 

MHA medium. Six 8-mm-diameter wells (holes) 

were drilled. An ethanol-based reference 

solution (99 percent ethanol) was also placed 

into one well, and leaves extracts of a trial 

medication at various concentrations were 

poured into each well, using the sterile pipette. 

There was a 50 l volume for all of the test and 

standard solutions tested. For 18 to 24 hours, 

the Petri dishes were incubated in an incubator 

at 37°C. An aseptic area was used for all 

operations. [8]Results: 

Vitex negundo's ethanolic extract was shown to 

have an antibacterial effect beginning at a 

concentration of 20mg/ml for all three 

microorganisms studied in this investigation 

(Fig-1). E. coli was shown to have antibacterial 

activity at all concentrations, with the greatest 

ZOI of 12 mm at 100 mg/ml (Table-1). At 80 

mg/ml and 100 mg/ml, the greatest zone of 

inhibition of 15 mm was obtained against S. 

aureus (Table-2). At a concentration of 

100mg/ml, Klebsiella pneumonia had the 

greatest ZOI of 11 mm (Table-3). 

 

 

 

 

 

 

 

 

Fig.1: Inhibition zone of Vitex negundo Linn. 

leaves extract 

 

Escherichia coli                          

Staphylococcus aureus              

Klebsiella pneumonia                          

Discussion: 



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The plant's active principle, in millimetres. 

Inverse polarity 

 

Gram-positive (S aureus) and gram-negative (E. 

coli) bacteria were both killed by the plant 

extract (E. coli, K. pneumoniae). A control 

medication of 98 percent ethanol was utilised. 

Due to the little amount (50 l) and high 

concentration of the control medication, it 

demonstrated minimal inhibition. Gram 

negative bacteria were less responsive to the 

experimental medication compared to gramme 

positive bacteria. Gram-negative bacteria tend 

to be more resistant to antibiotics than their 

gram-positive counterparts. [9] 

 

S. aureus was shown to have the widest area of 

inhibition, which is impressive given the 

organism's significance. There are a slew of 

things that might influence Antimicrobial 

properties are largely influenced by the 

extraction solvent. It may be possible that plant 

extracts include broad-spectrum antibiotics or 

just general metabolic poisons in them if they 

are able to kill both Gram positive and Gram 

negative bacteria. [10,11] 

 

Vitex negundo leaves and bark have been 

shown to have antibacterial and phytochemical 

properties in a prior research. 

 

Staphylococcus epidermidis, Bacillus subtilis, 

Staphylococcus aureus, and S. aureus were the 

three gram-positive bacteria tested, while E. 

coli, Salmonella typhimurium, Pseudomonas 

aeruginosa, and Vibrio anguillarum were the 

four gram-negative bacteria. Bacteria such as 

Cholera and Vibrion alginolyteus polar and non-

polar substanc  

  

Disc diffusion, Agar Cup and Broth Dilution 

procedures were used to test the antibacterial 

activity of the various extracts, including 

petroleum ether, chloroform, ethanol, and 

methanol. E. coli and S. aureus were shown to 

be the most resistant bacteria to extracts from 

both bark and leaves. Alcoholic leaf extracts 

inhibited the growth of bacteria in the positive 

and negative serogroups, but extracts from the 

tree bark in petroleum ether and chloroform 

exhibited superior antibacterial action. [12] 

Based on these and other studies, the trial 

medication seems to be an effective 

antibacterial therapeutic agent.. 

Conclusion                                                          

Antibacterial activity was shown to be present 

against all of the bacterial pathogens 

investigated in this investigation. Because of 

this, it may be employed as an antibacterial 

supplement as well as a potential novel 

medicinal drug. 

References: 

9. Infection Control in South-East Asia, 

World Health Organization South-East Asia 

Regional Office, New Delhi, Anonymous. In the 

year [2005, on the Internet], Accessed at: 

http://www.searo.who.int /entity/emerging 

diseases/documents/b0005.pdf [Cited on 5-9-

2016]. 

10. Second, Fauci AS. Infectious illnesses in 

the 21st century: considerations. Clinical 

Infectious Dis 

11.As an antibacterial agent, plant extracts and 

phytochemicals have been shown to be 

effective against antibiotic-resistant bacteria (4) 

13. 4th Purushottamacharyulu of Ganjam 

These are the fundamental ideas behind 

15. Ancient Indian Bacteriology and 

Ayurveda:Parekh J, Chanda S, Antibacterial 

activity of aqueous and methanol extracts of 



CTMJ | traditionalmedicinejournals.com                                    Chinese Traditional Medicine Journal | 2021 | Vol4 |Issue3 

 
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medicinal herbs. Turkey's Journal of Biology, 

Vol. 29, pp. 203-210 Medicinal herbs from 

South Africa have been shown to have 

antibacterial properties. Ethanopharmacol. 

1997;56(1):81-87 18. Antibiotics that are 

resistant to resistance. Scientific American. 

1995;270(5237):724-7. 

17. In the journal of medicinal plants 

research, Dutta and Panda report on the 

antibacterial activity and phytochemical 

screening of the leaf and bark extracts of Vitex 

negundo from the similipal biosphere reserve in 

Orissa. 

19.Patna, Shree, 1st ed., chapter 14 on 

literatureHow to cite the article: 

  

p.104-6 Baidyanath Ayurved Bhavan 

Commentary by Gorakhanath Chaturvedi and 

Kashinath Pandey, Reprint ed. 1p. 82 

5. Commentary on Dalhana's translation 

of the Sushrut Samhita, Sutra Sthana, Chapter 

38, Verse 18, is included in Yadavaji Trikamji's 

edition of the book, which was reprinted in 

Varanasi, India, in 2009. CCRAS; 2005;(3), page 

451 in Sharma P.C. et alDatabase .'s Medicinal 

Plants Used in Ayurveda Microbes in Action, by 

Seely H.W. and P.J. Van Denmark. The 2nd 

edition of A Laboratory Manual of Microbiology. 

It was published in Bombay, India, in 1975 by D 

B Taraporevala Sons & Co Pvt Ltd.  

Kethamakka Shivarama Prasad, Kunal Kale, 

Meena Shamrao Deogade and Nilima 

Tankhiwale are the other members of the team. 

Vitex Negundo Linn. (Nirgundi) Leaves Extract 

Antimicrobial Activity. Trad. Med. Research 

2016, 2(4): 99-102. 

http:dx.doi.org/10.21276/jrtm.2016/206 

Source of Support: 

Datta Meghe Institute of Medical Sciences (DU), 

Nagpur 

Con ict of Interest: Nil 

 

© Journal of Research in Traditional Medicine 

2015-2016 

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resulting from the use of the information 

included in published articles, neither Journal of 

Research in Traditional Medicine, its publisher, 

editorial board members, nor anyone else 

involved in their production or delivery, 

assumes any duty or obligation. In the pieces, 

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